PKC gamma CRISPR Activation Plasmid (h): sc-400503-ACT

PRKCG encodes protein kinase C gamma (PKCγ), a conventional Ca2+/diacylglycerol-dependent serine/threonine kinase enriched in neuronal tissues that transduces signals downstream of phospholipase C and receptor-mediated phosphoinositide turnover. PKCγ phosphorylates diverse substrates to modulate synaptic signaling, cytoskeletal dynamics, and activity-dependent gene expression, integrating second-messenger cues into MAPK/ERK and other kinase networks. In the central nervous system, PRKCG activity influences neurotransmission and neuronal plasticity, and altered regulation has been associated with neurodegenerative and neurodevelopmental phenotypes, including hereditary ataxias. These properties make PRKCG a useful node for dissecting signal transduction mechanisms and pathway cross-talk in human cellular and neuronal models.

PKC gamma CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKCG expression without altering the underlying DNA sequence.

PKC gamma CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKCG locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PRKCG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKC gamma expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKCG locus and enabling the study of PKC gamma-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKC gamma pathway restoration in tumor cells with silenced or reduced PRKCG expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.